35th Annual Student Research Conference
Investigation into the Efficacy of Urea-Spiked Lysozyme Assays to Demonstrate the Chaperone Activity of DBF Enzyme on Protein Aggregates
Guthrie L. Stroh
Dr. Cassidy Dobson, Faculty Mentor
Disulfide Bond-Forming enzyme (DBF) is a small molecular chaperone protein isolated from the extremophilic archaebacterium Sulfolobus solfataricus. DBF has the unique ability to rearrange disulfide bonds without itself containing the cysteine residue(s) quintessential to similarly functioning chaperones (e.g. PDI). In order to elucidate this idiosyncratic structure, it is first necessary to study the effectiveness of DBF to refold protein aggregates (incorrectly folded clusters held together by disulfide bonds). Protein aggregates are causal of many diseases such as cataracts and Alzheimer’s, so a protein like DBF could have significant medical application if proven effective. DBF’s activity will be studied through the use of the lysozyme assay, which will test the ability of DBF to refold denatured lysozyme under various environmental conditions in vitro through UV-Vis spectrophotometry. Previous work has shown the chaotropic agent urea to be a potential assay additive that could assist in retaining aggregates in solution during treatment with DBF. The present work will therefore investigate the applicability of urea to the lysozyme assay’s ability to demonstrate the functionality of DBF.
Keywords: Disulfide Bond-Forming enzyme (DBF), protein aggregates, lysozyme assay, molecular chaperones, urea
Topic(s):Biochemistry and Molecular Biology
Presentation Type: Oral Presentation
Session: 105-5
Location: MG 2001
Time: 9:30